Laboratory of Crop Physiology, Research Faculty of Agriculture, Hokkaido University Kita 9 Nishi 9 Kita-ku, Sapporo-shi, Hokkaido, 060-8589, Japan; Asian Natural Environmental Science Center, The University of Tokyo 1-1-1 Midori-cho, Nishitokyo-shi, Tokyo, 188-0002, Japan.
State Key Laboratory of Subtropical Silviculture, Zhejiang A & F University, Lin'an, Hangzhou, 311300, PR China.
Biochem Biophys Res Commun. 2018 Oct 20;505(1):176-180. doi: 10.1016/j.bbrc.2018.09.088. Epub 2018 Sep 20.
Protein phosphatase 2A (PP2A) is a heterotrimeric protein complex conserved among eukaryotes. The B subunit of PP2A determines the substrate specificity of the PP2A holoenzyme, and is classified into the B, B', B″ and B‴ families. Arabidopsis thaliana has two isoforms of the B-family subunit (ATBA and ATBB). A double knockout of their genes is lethal, but which developmental process is primarily impaired by the double knockout is unclear. Identifying such a process helps understand PP2A-mediated signaling more deeply. Here, genetic characterization of new knockout mutants for these genes shows that they are necessary for pollen development but not for female gametophyte development. Compared to wild-type pollen grains, the mutant pollen grains exhibited lower enzyme activities, germinated less frequently on stigmas, and exhibited the aberrant numbers of sperm cell nuclei, suggesting that ATBA and ATBB play pleiotropic roles in pollen development. The amino acids stabilizing the interaction between the human PP2A A and B-family subunits are conserved in an Arabidopsis A subunit (AtPP2AA2), ATBA and ATBB. His-tagged AtPP2AA2 co-immunoprecipitated with either Myc-tagged ATBA or Myc-tagged ATBB in vitro, confirming their interactions. Proteins that regulate pollen development and that undergo dephosphorylation are likely primary targets of ATBA and ATBB.
蛋白磷酸酶 2A(PP2A)是一种在真核生物中保守的异三聚体蛋白复合物。PP2A 的 B 亚基决定了 PP2A 全酶的底物特异性,并分为 B、B'、B″和 B‴家族。拟南芥有两种 B 家族亚基(ATBA 和 ATBB)的同工型。它们的基因双敲除是致死的,但双敲除主要影响哪个发育过程尚不清楚。鉴定这样的过程有助于更深入地了解 PP2A 介导的信号转导。在这里,对这些基因的新敲除突变体的遗传特征表明,它们对于花粉发育是必需的,但对于雌性配子体发育不是必需的。与野生型花粉粒相比,突变型花粉粒的酶活性较低,在柱头萌发的频率较低,并且精子核的数量异常,表明 ATBA 和 ATBB 在花粉发育中发挥多效性作用。稳定人 PP2A A 和 B 家族亚基相互作用的氨基酸在拟南芥 A 亚基(AtPP2AA2)、ATBA 和 ATBB 中保守。体外 His 标记的 AtPP2AA2 与 Myc 标记的 ATBA 或 Myc 标记的 ATBB 共免疫沉淀,证实了它们的相互作用。调节花粉发育并经历去磷酸化的蛋白质可能是 ATBA 和 ATBB 的主要靶标。